Under anaerobic conditions, Saccharomyces cerevisiae uses NADH-dependent glycerol-3-phosphate dehydrogenase (Gpd1p and Gpd2p) to re-oxidize excess NADH, yielding substantial amounts of glycerol. In a Deltagpd1 Deltagpd2 double-null mutant, the necessary NAD(+) regeneration through glycerol production is no longer possible, and this mutant does not grow under anaerobic conditions. The excess NADH formed can potentially be used to drive other NADH-dependent reactions or pathways. To investigate this possibility, a double-null mutant was transformed with a heterologous gene (mt1D) from Escherichia coli, coding for NADH-dependent mannitol-I-phosphate dehydrogenase. Expression of this gene in S. cerevisiae should result in NADH oxidation by the NADH-requiring formation of mannitol-1-phosphate from fructose-6-phosphate. The strain was characterized using step-change experiments, in which, during the exponential growth phase, the inlet gas was changed from air to nitrogen. It was found that the mutant produced mannitol only under anaerobic conditions. However, anaerobic growth was not regained, which was probably due to the excessive accumulation of mannitol in the cells. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

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BibTeX @article{Costenoble2003,author={Costenoble, R. and Adler, Lennart and Niklasson, Claes and Liden, G.},title={Engineering of the metabolism of Saccharomyces cerevisiae for anaerobic production of mannitol},journal={Fems Yeast Research},issn={1567-1356},volume={3},issue={1},pages={17-25},abstract={Under anaerobic conditions, Saccharomyces cerevisiae uses NADH-dependent glycerol-3-phosphate dehydrogenase (Gpd1p and Gpd2p) to re-oxidize excess NADH, yielding substantial amounts of glycerol. In a Deltagpd1 Deltagpd2 double-null mutant, the necessary NAD(+) regeneration through glycerol production is no longer possible, and this mutant does not grow under anaerobic conditions. The excess NADH formed can potentially be used to drive other NADH-dependent reactions or pathways. To investigate this possibility, a double-null mutant was transformed with a heterologous gene (mt1D) from Escherichia coli, coding for NADH-dependent mannitol-I-phosphate dehydrogenase. Expression of this gene in S. cerevisiae should result in NADH oxidation by the NADH-requiring formation of mannitol-1-phosphate from fructose-6-phosphate. The strain was characterized using step-change experiments, in which, during the exponential growth phase, the inlet gas was changed from air to nitrogen. It was found that the mutant produced mannitol only under anaerobic conditions. However, anaerobic growth was not regained, which was probably due to the excessive accumulation of mannitol in the cells. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.},year={2003},keywords={delta gpd1 delta gpd2 double-null mutant, redox balance, nadh-coupled reduction, mannitol-1-phosphate dehydrogenase, glycerol formation, glycerol-3-phosphate dehydrogenase mutants, escherichia-coli, nadph production, osmotic-stress, glycerol, yeast, cultures, growth, gpd1, gene },}

RefWorks RT Journal ArticleSR ElectronicID 40668A1 Costenoble, R.A1 Adler, LennartA1 Niklasson, ClaesA1 Liden, G.T1 Engineering of the metabolism of Saccharomyces cerevisiae for anaerobic production of mannitolYR 2003JF Fems Yeast ResearchSN 1567-1356VO 3IS 1SP 17OP 25AB Under anaerobic conditions, Saccharomyces cerevisiae uses NADH-dependent glycerol-3-phosphate dehydrogenase (Gpd1p and Gpd2p) to re-oxidize excess NADH, yielding substantial amounts of glycerol. In a Deltagpd1 Deltagpd2 double-null mutant, the necessary NAD(+) regeneration through glycerol production is no longer possible, and this mutant does not grow under anaerobic conditions. The excess NADH formed can potentially be used to drive other NADH-dependent reactions or pathways. To investigate this possibility, a double-null mutant was transformed with a heterologous gene (mt1D) from Escherichia coli, coding for NADH-dependent mannitol-I-phosphate dehydrogenase. Expression of this gene in S. cerevisiae should result in NADH oxidation by the NADH-requiring formation of mannitol-1-phosphate from fructose-6-phosphate. The strain was characterized using step-change experiments, in which, during the exponential growth phase, the inlet gas was changed from air to nitrogen. It was found that the mutant produced mannitol only under anaerobic conditions. However, anaerobic growth was not regained, which was probably due to the excessive accumulation of mannitol in the cells. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.LA engPMID 12702242DO 10.1111/j.1567-1364.2003.tb00134.xLK http://dx.doi.org/10.1111/j.1567-1364.2003.tb00134.xOL 30